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目的:探讨雌激素相关受体α(estrogen-related receptorα,ERRα)与雌激素受体α(estrogen receptors alpha,ERα)经典通路的关系。方法:采用蛋白质印迹法观察雌二醇(E2)、ICI182、780及XCT790对MCF-7细胞中ERRα、ERα和TFF1蛋白表达的影响;Real-time RT-PCR检测MCF-7细胞中ERRα、ERα和TFF1mRNA表达水平的变化。结果:E2诱导后ERα由(100.0±0.0)%增加至(240.0±27.0)%,ERRα由(100.0±0.0)%增加至(244.2±35.9)%,TFF1mRNA由(100.0±0.0)%增加至(201.7±21.6)%;ERα蛋白的表达由(100.0±0.0)%增加至(128.4±5.3)%,ERRα蛋白的表达由(100.0±0.0)%增加至(157.3±5.6)%,TFF1蛋白的表达由(100.0±0.0)%增加至(133.2±7.1)%,表明E2可上调这3种基因的表达。ICI182、780作用后ERα、ERRα和TFF1mRNA表达量分别为(133.6±6.9)%、(168.1±26.8)%和(138.9±12.9)%,蛋白表达量分别为(84.2±5.0)%、(100.0±7.5)%和(84.9±5.7)%,表明ICI182、780可抑制E2对ERRα、ERα和TFF1的诱导作用。XCT790作用后ERRα和TFF1 mRNA表达量分别为(136.8±28.2)%和(140.4±11.4)%,ERα、ERRα和TFF1蛋白表达量分别为(76.4±5.3)%、(90.7±6.9)%和(76.3±6.6)%,表明ICI182、780和XCT790均对E2诱导的ERα、ERRα、TFF1 mRNA和蛋白上调有抑制作用。ICI182、780与XCT790联合作用后,ERα、ERRα和TFF1mRNA表达量分别为(98.4±17.2)%、(108.7±8.8)%和(95.1±21.6)%,蛋白表达量分别为(58.2±6.7)%、(63.2±6.0)%和(57.1±5.9)%,表明两种阻断剂联合作用对这种上调的抑制作用大于二者单独的抑制作用。结论:E2对ERRα的诱导作用是通过ER介导的,将ERα和ERRα联合应用作为分子生物学指标及靶标,对乳腺癌内分泌治疗策略的实施可能有其临床意义。
Objective: To investigate the relationship between estrogen-related receptor α (ERRα) and estrogen receptor alpha (ERα). Methods: The effects of estradiol (E2), ICI182,780 and XCT790 on the expression of ERRα, ERα and TFF1 protein in MCF-7 cells were observed by Western blotting. The expressions of ERRα, ERα And TFF1 mRNA expression level changes. Results: The ERα increased from (100.0 ± 0.0)% to (240.0 ± 27.0)%, the ERRα increased from (100.0 ± 0.0)% to (244.2 ± 35.9)% and the TFF1 mRNA increased from (100.0 ± 0.0)% to 201.7 ± 21.6%). The expression of ERαprotein increased from (100.0 ± 0.0)% to (128.4 ± 5.3)% and the expression of ERRαprotein increased from (100.0 ± 0.0)% to (157.3 ± 5.6)% Increased from (100.0 ± 0.0)% to (133.2 ± 7.1)%, indicating that E2 can up-regulate the expression of these three genes. The expressions of ERα, ERRα and TFF1mRNA were (133.6 ± 6.9)%, (168.1 ± 26.8)% and (138.9 ± 12.9)% respectively after ICI182,780 treatment and the protein expression levels were (84.2 ± 5.0)% and 7.5)% and (84.9 ± 5.7)%, respectively, indicating that ICI182,780 inhibits the induction of ERRα, ERα and TFF1 by E2. The expression of ERRα and TFF1 mRNA were (136.8 ± 28.2)% and (140.4 ± 11.4)%, respectively, and the expression of ERα, ERRα and TFF1 protein were (76.4 ± 5.3)% and (90.7 ± 6.9)% 76.3 ± 6.6)%, indicating that ICI182,780 and XCT790 both inhibited the up-regulation of E2 induced ERα, ERRα, TFF1 mRNA and protein. The expression of ERα, ERRα and TFF1 mRNA were (98.4 ± 17.2)%, (108.7 ± 8.8)% and (95.1 ± 21.6)% respectively after ICI182,780 combined with XCT790. The expression of ERα, ERRα and TFF1 mRNA were 58.2 ± 6.7% , (63.2 ± 6.0)% and (57.1 ± 5.9)%, respectively, indicating that the combined effect of the two blockers inhibited the up-regulation more than the separate inhibition. CONCLUSION: The induction of ERRα by E2 is mediated by ER. Combined use of ERα and ERRα as a molecular biological target and target may have clinical significance for the implementation of breast cancer endocrine therapy strategy.